Affiliation:
1. Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical, Lima, Perú
2. Centre de Recerca en Salut Internacional de Barcelona, Hospital Clinic/Institut d'Investigacions Biomèdiques August Pi i Sunyer, Universitat de Barcelona, Barcelona, Spain
3. CIBERESP, Barcelona, Spain
4. Department of Epidemiology, University of Texas School of Public Health, Houston, Texas, USA
Abstract
ABSTRACT
Infectious diarrhea can be classified based on its clinical presentation as noninflammatory or inflammatory disease. In developing countries, among inflammatory diarrhea cases,
Shigella
is the most common cause, followed by
Campylobacter
and
Salmonella
. Because the time frame in which treatment choices must be made is short and conventional stool cultures lack good sensitivity, there is a need for a rapid, sensitive, and inexpensive detection technique. The purpose of our study was to develop a multiplex real-time PCR procedure to simultaneously identify
Campylobacter
spp.,
Salmonella
spp., and
Shigella
spp. Primers were designed to amplify the
invA
,
ipaH
, and 16S rRNA genes simultaneously in a single reaction to detect
Salmonella
,
Shigella
, and
Campylobacter
, respectively. Using this approach, we correctly identified 102 of 103 strains of the targeted enteropathogens and 34 of 34 other pathogens. The melting temperatures were 82.96 ± 0.05°C for
invA
, 85.56 ± 0.28°C for
ipaH
, and 89.21 ± 0.24°C for 16S rRNA. The limit of accurate quantification for the assay in stool samples was 10
4
CFU g
−1
; however, the limit of detection was 10
3
CFU g
−1
. This assay is a simple, rapid, inexpensive, and reliable system for the practical detection of these three enteropathogens in clinical specimens.
Publisher
American Society for Microbiology
Cited by
46 articles.
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